不同低氧浓度对人脐带间充质干细胞向神经细胞分化的影响  

Effects of Different Hypoxia Concentrations on Differentiation of Human Umbilical Cord Mesenchymal Stem Cells into Neurons

作　　者：罗芸 马俊[2,3] 钱前 张富婷 王晓辉[2,3] LUO Yun;MA Jun;QIAN Qian;ZHANG Fu-ting;WANG Xiao-hui(The Third Department of Cadre Wards,940 Hospital of PLA Joint Logistics Support Forces,Lanzhou 730050,China;Department of Medical Experiment,940 Hospital of PLA Joint Logistics Support Forces,Lanzhou 730050,China;Key Lab of Stem Cells and Genetaceutical of Gansu Province)

机构地区：[1]中国人民解放军联勤保障部队第九四O医院干部病房三科,兰州730050 [2]中国人民解放军联勤保障部队第九四O医院医学实验科,兰州730050 [3]中国人民解放军联勤保障部队第九四O医院甘肃省干细胞与基因药物重点实验室

出　　处：《解放军医药杂志》2019年第8期6-11,共6页Medical & Pharmaceutical Journal of Chinese People’s Liberation Army

基　　金：全军医药卫生科研基金课题(CLZ15JA06)

摘　　要：目的探讨不同低氧浓度对人脐带间充质干细胞(hUCMSCs)诱导分化成神经细胞的促进作用。方法hUCMSCs分为对照组(常氧浓度)和实验组(5%氧浓度组、10%氧浓度组)。MTT法检测细胞增殖;乳酸脱氢酶(LDH)法检测细胞损伤;流式细胞仪检测细胞凋亡;WestonBlot检测细胞神经生长因子(NGF)、神经巢蛋白(Nestin)、神经元特异性烯醇化酶(NSE)的表达。结果hUCMSCs细胞在缺氧条件下细胞增殖受到抑制,培养24h内实验组OD值显著低于对照组(P<0.05),随着培养时间的延长,实验组细胞增殖加快,到培养至96h时实验组与对照组比较差异无统计学意义(P>0.05)。在培养24、48h时,实验组hUCMSCs细胞LDH活性显著高于对照组(P<0.05);培养至72h后5%氧浓度组LDH活性显著高于对照组(P<0.05),10%氧浓度组与对照组比较差异无统计学意义(P>0.05);培养至96h后各组hUCMSCs细胞LDH的活性比较差异无统计学意义(P>0.05)。5%氧浓度组hUCMSCs细胞抗凋亡作用强于10%氧浓度组(P<0.05)。缺氧培养的hUCMSCs细胞更早出现神经元样形态学变化,且细胞中NGF、Nestin、NSE的表达水平高于对照组(P<0.05)。结论5%氧浓度缺氧培养的hUCMSCs细胞具有抗细胞凋亡作用,同时对成神经诱导分化具有促进作用。Objective To investigate effects of different hypoxia concentrations on differentiation of human umbilical cord mesenchymal stem cells(hUCMSCs)into neurons.Methods The hUCMSCs were divided into control group(normoxia concentration)and experimental groups(5%oxygen concentration group and 10%oxygen concentration group).MTT assay was used to detect cell proliferation;lactate dehydrogenase(LDH)assay was used to detect cell damage;flow cytometry was used to detect cell apoptosis;Weston Blot method was used to detect expressions of nerve growth factor(NGF),neural nest(Nestin)and neuron specific enolase(NSE).Results The hUCMSCs proliferation was inhibited under hypoxia condition.Values of optical density difference(OD)in experimental groups were significantly lower than that in control group within 24 h of culture(P<0.05);hUCMSCs proliferation in experimental groups were accelerated with prolongation of culture time;there were no significant differences in hUCMSCs proliferation between experimental groups with control group in 96 h of culture(P>0.05).LDH activities of hUCMSCs in experimental groups were significantly higher than those in control group in 24 h and 48 h of culture(P<0.05).LDH activity in 5%oxygen concentration group was significantly higher than that in control group in 72 h of culture(P<0.05),but there was no significant difference in LDH activity between 10%oxygen concentration group with control group(P>0.05).There were no significant differences in LDH activity of hUCMSCs among groups in 96 h of culture(P>0.05).The anti-apoptotic effect in 5%oxygen concentration group was better than that in 10%oxygen concentration group(P<0.05).Neuron-like morphological changes were earlier appearance in hypoxic cultured hUCMSCs,and NGF,Nestin and NSE expressions in hypoxic cultured hUCMSCs were significantly higher than those in control group(P<0.05).Conclusion hUCMSCs cultured in 5%oxygen concentration under hypoxia condition have anti-apoptotic effect and promoted neurogenesis.

关 键 词：低氧 人脐带间充质干细胞 细胞分化 神经细胞 

分 类 号：R392.2[医药卫生—免疫学] R594.3[医药卫生—基础医学]